Figure 5
From: The Na/K-ATPase α1 and c-Src form signaling complex under native condition: A crosslinking approach
Immunoblotting analysis for the α1 subunit, c-Src, and α1 + c-Src in LLC-PK1 cells by capillary immunoblotting: Preparation of whole cell lysate with Native-PAGE sample buffer were performed as described for BN-PAGE in the Materials and Methods. Same DTT/SDS–treated, BMH-crosslinked samples (3 μg protein/sample) of LLC-PK1 cells were electrophoresed side-by-side on the same 66–440 kDa separation module and immunoblotted with detection module (Wes system, ProteinSimple) with antibodies against the α1 (upper panel), c-Src (middle panel), and α1 + c-Src (lower panel, mixture of the two antibodies with same dilutions for α1 alone and c-Src alone) separately. For DTT/SDS treatment, samples were heated at 60 °C for 30 min (for the α1 subunit and α1 + c-Src) or 95 °C for 5 min (for c-Src). Please note there are multiple peaks. n = 3.
